Pharmaceutical composition for preventing or treating parkinson&#39;s disease, comprising 2-(4-(1-hydroxypropane-2-yl)phenyl)isoindoline-1-one compound

ABSTRACT

The present invention relates to a pharmaceutical composition for preventing or treating Parkinson&#39;s disease, comprising a 2-(4-(1-hydroxypropane-2-yl)phenyl)isoindoline-1-one compound or a pharmaceutically acceptable salt thereof, wherein the pharmaceutical composition for preventing or treating Parkinson&#39;s disease can increase the protein level of PGC-1α in the brain of an individual by successfully passing through the blood-brain barrier (BBB).

TECHNICAL FIELD

The present disclosure relates to a pharmaceutical composition forpreventing or treating Parkinson's disease, including a2-(4-(1-hydroxypropan-2-yl)phenyl)isoindolin-1-one compound or apharmaceutically acceptable salt thereof.

BACKGROUND ART

Parkinson's disease, a disease whose main symptoms are tremors(shaking), rigidity, ataxia (slowed movements), and prolonged unstableposture, is a chronic disease caused by a lack of neurotransmitterscalled dopamine in the brain and one of the degenerative diseases in thecentral nervous system, which is started with modification in thesubstantia nigra pars compacta in the midbrain and accompanied by itspathophysiological symptoms such as reduction in brain volume andaggregation of α-synuclein (αSyn) as well as imperfect gait, handtremor, and rigid behavior.

Most treatment strategies for Parkinson's disease were limited tomanaging the symptoms of motor functions with drugs such as L-DOPA ordopamine receptor agonists as well as deep brain stimulation. Inaddition, these therapies at the current research level have failed toprevent the gradual death of dopaminergic neurons (DAs).

Recently, on the other hand, when it comes to death and survival ofcells, research related to the function of peroxisomeproliferator-activated receptor-γ coactivator-1α (PGC-1α) and variousdiseases that may be caused by dysregulation of PGC-1α has beenreported.

Neurodegenerative diseases such as Alzheimer's disease, Parkinson'sdisease, Huntington's disease, and Lou Gehrig's disease are caused bythe gradual loss of function and death of neurons, and the overallsymptoms of these diseases are due to the loss of certain parts ofneurons. Unlike the hyperactivity due to neurodegeneration observed inPGC-1α knock-out mice and the damaged areas that are less observed inthe cerebral cortex, it is noticed that PGC-1α is directly related toneurodegenerative diseases based on the damaged sites that are markedlyfound in corpus striatum in the brain.

Also, in addition to these findings, the identification of vacuolarlesions in the central nervous system in PGC-1α knock-out mice showsthat PGC-1α plays a crucial role in maintaining neuronal functions.

Reduced expression of PGC-1α increases the expression of BACE1, whichproduces beta-amyloid by degrading and cleaving the progenitor proteinof amyloid that causes Alzheimer's disease and increases the amount ofbeta-amyloid, thereby leading to mitochondrial hypofunction and celldeath. Single-nucleotide mutations in the PGC-1α gene are highlycorrelated with an increase in the risk factors that develop Parkinson'sdisease and Huntington's disease and are known to decrease expression ofPGC-1α genes in patients with Alzheimer's disease, Parkinson's disease,and Huntington's disease.

Recently, there has been a rising interest in treatment methods forParkinson's disease by pharmacologically activating PGC-1α for itsfunctional mechanisms, which are closely related to thoseneurodegenerative diseases.

PRIOR ART DOCUMENT Patent Document

(Patent Document 1) Korean Patent No. 10-1384642

DISCLOSURE OF THE INVENTION Technical Goals

An object of the present disclosure is to provide a composition capableof preventing or treating Parkinson's disease in a subject, as anexpression of PGC-1α is enhanced in the brain by including a2-(4-(1-hydroxypropan-2-yl)phenyl)isoindoline-1-one compound.

Technical Solutions

The present disclosure provides a pharmaceutical composition forpreventing or treating Parkinson's disease, including a compoundrepresented by the following Chemical Formula 1 or a pharmaceuticallyacceptable salt thereof as an active ingredient.

In addition, the composition may increase the expression of peroxisomeproliferator-activated receptor-γ coactivator-1α (PGC-1α).

In addition, the composition may have a formulation selected from thegroup consisting of a solution, suspension, syrup, emulsion, liposome,acid, powder, granule, tablet, sustained-release agent, and capsule.

In addition, the composition may be a composition for oraladministration and have a formulation of a drug carrier orsustained-release agent, including liposomes.

In addition, the composition may be a composition for parenteraladministration and have a formulation of a drug carrier orsustained-release agent including liposomes and ultrasonic contrastagents.

In addition, the compound represented by Chemical Formula 1 is2-(4-(1-hydroxypropan-2-yl)phenyl)isoindoline-1-one.

According to another aspect of the present disclosure, healthyfunctional food for alleviating Parkinson's disease, including acompound represented by the Chemical Formula 1 as an active ingredient,is provided.

According to another aspect of the present disclosure, a method ofpreventing or treating Parkinson's disease, including administering acomposition including a compound represented by the Chemical Formula 1or a pharmaceutically acceptable salt thereof, to an individual may beprovided.

Advantageous Effects

The pharmaceutical composition for preventing or treating Parkinson'sdisease, according to the present disclosure, successfully passesthrough the blood-brain barrier (BBB) and increases expression ofPGC-1α, which has a neuroprotective ability in the brain of anindividual, thereby exhibiting the effect of preventing or treatingParkinson's disease of a subject.

BRIEF DESCRIPTION OF DRAWINGS

FIG. 1 shows a reaction scheme illustrating the synthetic process of a2-(4-(1-hydroxypropan-2-yl)phenyl)isoindolin-1-one compound according tothe present disclosure.

FIG. 2 shows NMR data for identifying a structure after synthesizing a2-(4-(1-hydroxypropan-2-yl)phenyl)isoindolin-1-one compound according tothe present disclosure.

FIG. 3 shows a schematized compound screening experimental method inExample 2.

FIG. 4 shows an activity measurement result graph for the PGC-1αpromoter via luciferase assay in Example 2.

FIG. 5 shows a graph illustrating the result of experiments performedwith 14 drugs that increase the activity of the PGC-1α promoter by 2.5times or more.

FIG. 6 shows an image of proteins from an immunoblot after treating anSH-SY5Y cell line with compounds (Example 4).

FIG. 7 shows an immunoblot of PGC-1α protein in the substantia nigra(SN) of mice fed with compounds (Example 4).

FIG. 8 shows immunohistochemistry and immunoblot in the brain tissue ofParkinson's disease model mice fed with a compound-included diet(Example 5).

FIG. 9 shows a pole test of Parkinson's disease model mice fed with acompound-included diet (Example 6).

FIG. 10 shows the expression of PGC-1α and the primary target genes ofPGC-la in Parkinson's disease model mice fed with a compound-includeddiet (Example 7).

BEST MODE FOR CARRYING OUT THE INVENTION

Since the present disclosure may be subjected to various modificationsand have various example embodiments, specific example embodiments willbe illustrated in the drawings and described in detail. However, this isnot intended to limit the present disclosure to specific exampleembodiments and should be understood to include all modifications,equivalents, and substitutes included in the spirit and scope of thepresent disclosure. In describing the present disclosure, when it isdetermined that the detailed description of the related known techniquesmay obscure the gist of the present disclosure, the detailed descriptionthereof will be omitted.

The present disclosure provides a pharmaceutical composition forpreventing or treating Parkinson's disease, including a compoundrepresented by the following Chemical Formula 1 or a pharmaceuticallyacceptable salt thereof as an active ingredient.

Hereinafter, the pharmaceutical composition for preventing or treatingParkinson's disease according to a specific embodiment of the presentdisclosure will be described in more detail.

Conventional treatment strategies for Parkinson's disease have beenlimited to managing symptoms of motor functions using drugs such asL-DOPA or dopamine receptor agonists and deep brain stimulation, whichhave failed to prevent the gradual death of dopaminergic neurons (DAs).

Accordingly, the present inventors completed the present disclosure byidentifying that when a specific derivative compound is administered,the compound successfully passes through the BBB to increase theactivity of the PGC-1α promoter as well as the level of proteinexpression of PGC-1α in the brain.

The present disclosure provides a pharmaceutical composition forpreventing or treating Parkinson's disease, including the2-(4-(1-hydroxypropan-2-yl)phenyl)isoindoline-1-one compound or apharmaceutically acceptable salt thereof as an active ingredient.

PGC-1α is a key regulator of mitochondrial function to co-regulatetranscriptional programs that are important for mitochondrial biogenesisand protect mitochondria from oxidative stress. Such PGC-1α leveldecreases in patients with Parkinson's disease, and the decline in thePGC-1α level in Parkinson's disease is considered to be due tomethylation on the PGC-1α promoter.

On the other hand, PGC-1α knock-out mice are more sensitive to thedegenerative effect of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP), a Parkinson's disease neurotoxin, overexpression of PGC-1α isknown to have a protective effect against N-methyl-4-phenylpyridiniumion (MPP+) toxin, an active metabolite of MPTP, and overexpression ofPGC-1α is also known to exhibit a protective effect against α-synuclein,MPTP, oxidative stress and rotenone-induced degeneration.

The PGC-1α reactive gene is downregulated in dopaminergic neuronsderived from patients with Parkinson's disease, which is thought tosuggest that the PGC-1α plays an important role in the cause ofParkinson's disease; and, when PARIS, a substrate of Parkin which is acausative protein of Parkinson's disease, is overexpressed, the loss ofdopamine neurons is suppressed by PGC-1α overexpression, which isthought to show that PGC-1α is the main target of Parkin in dopaminergicneurodegeneration.

Therefore, defects in PGC-1α signaling have emerged as an importantcause of dopaminergic degeneration in Parkinson's disease, and reductionof PGC-1α due to dysfunction of Parkin may be a prime target for theprevention or treatment of Parkinson's disease.

In Example 2 of the present disclosure, activity measurement results ofthe promoter of PGC-1α through a luciferase assay were shown (FIG. 4 ),and the results of experiments performed with 14 drugs that increase theactivity of PGC-1α promoter by 2.5 times or more were shown, wherein theactivity of 2-(4-(1-hydroxypropan-2-yl)phenyl)isoindoline-1-one which isa compound represented by Chemical Formula 1 was the highest (FIG. 5 ).

In addition, in Example 4 of the present disclosure, according to theimage analysis of immunoblot performed after treating an SH-SY5Y cellline with various concentrations of compounds (0, 0.01, 0.1, 1.0, 10 μM,48 hours), it was found that2-(4-(1-hydroxypropan-2-yl)phenyl)isoindoline-1-one increases the amountof PGC-1α protein even in the low concentrations (FIG. 6 ).

In addition, as shown in immunoblot for PGC-1α protein in the SN of micefed with Chow, Indoprofen, or YPD-01(2-(4-(1-hydroxypropan-2-yl)phenyl)isoindoline-1-one) diet (0.5% w/w),it was found that 2-(4-(1-hydroxypropan-2-yl)phenyl)isoindoline-1-onesuccessfully passed through the BBB and statistically significantlyincreased the protein expression of PGC-1α in the SN of mice (Example 4,FIG. 7 ).

In addition, in Example 5 of the present disclosure, after feedingParkinson's disease model mice with Chow or YPD-01 diet (0.5% w/w) andthen sectioning the brain using a microtome after collecting the brain,AAV-PARIS injection remarkably killed dopamine neurons that wassignificantly inhibited by the administration of YPD-01 diet (FIG. 8 ).

In addition, in Example 6 of the present disclosure, as a result ofperforming a pole test after feeding Parkinson's disease model mice withChow or YPD-01 diet (0.5% w/w), the time required forAAV-PARIS8-injected mice to come down from the top of the pole tooktwice longer than AAV-GFP-injected mice, and it was found thatbehavioral abnormality disappeared due to YPD-01 administration (FIG. 9).

In addition, in Example 7 of the present disclosure, as a result ofmeasuring the expression of PGC-1α and the target genes of PGC-1α afterfeeding Parkinson's disease model mice with Chow or YPD-01 diet (0.5%w/w), over-expression of PARIS by AAV-PARIS brought about suppression ofPGC-1α expression and reduced expression of target genes thereof. It wasfound that the suppression of expression of PGC-1α and main target genesof PGC-1α (NRF-1, Tfam) by PARIS in the substantia nigra of mice fedwith YPD-01 diet was significantly restored (FIG. 10 ). [52].

On the other hand, the compound represented by the following ChemicalFormula 1 is 2-(4-(1-hydroxypropan-2-yl)phenyl)isoindoline-1-one.

FIG. 1 is a synthesis reaction scheme of the2-(4-(1-hydroxypropan-2-yl)phenyl)isoindoline-1-one compound, and FIG. 2is NMR data of identifying a structure after synthesizing the compound.

In the pharmaceutical composition of the present disclosure, the activeingredient is a compound of Chemical Formula I, a pharmaceuticallyacceptable salt thereof, a hydrate, or a solvate.

The term “pharmaceutically acceptable salt” as used herein refers to asalt of the compound that induces the desired pharmacological effect,that is, the expression of PGC-1α. Such salts may be formed by usinginorganic acids such as hydrochloride, hydrobromide, and hydroiodide, aswell as organic acid such as acetate, adipate, alginate, aspartate,benzoate, benzene sulfate, p-toluene sulfonate, bisulfate, sulfamate,sulfate, naphthylate, butyrate, citrate, camporate, camposulfonate,cyclopentane propionate, digluconate, dodecyl sulfate, ethane sulfonate,fumarate, glucoheptanoate, glycerophosphate, hemisulfate, heptanoate,hexanoate, 2-hydroxyethane sulfate, lactate, maliate, methane sulfonate,2-naphthalene sulfonate, nicotinate, oxalate, tosylate, and undecanoate.The term “pharmaceutically acceptable hydrate” as used herein refers toa hydrate of the compound having the desired pharmacological effect, andthe term “pharmaceutically acceptable solvate” as used herein refers toa solvate of the compound having the desired pharmacological effect. Thehydrate and solvate may also be prepared using the acid described above.

On the other hand, suitable carriers, excipients, and diluents that arecommonly used to prepare the pharmaceutical composition may further beincluded. In addition, it may be formulated according to conventionalmethods into oral formulations such as acids, granules, tablets,capsules, suspensions, emulsions, syrups, and aerosols, as well as formsof external agents, suppositories, and sterile injection solutions.

Carriers, excipients, and diluents that may be included in thecomposition are lactose, dextrose, sucrose, sorbitol, mannitol, xylitol,erythritol, maltitol, starch, gum acacia, alginate, gelatin, calciumphosphate, calcium silicate, cellulose, methylcellulose,microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propyl hydroxybenzoate, talc, magnesium stearate, andmineral oil. When the composition is formulated, it may be preparedusing diluents or excipients such as fillers, extenders, binders,wetting agents, disintegrants, surfactants, and the like that arecommonly used.

The pharmaceutical composition, according to the present disclosure, maybe administered in a pharmaceutically effective amount. The term“pharmaceutically effective amount” as used herein refers to an amountsufficient to treat a disease at a reasonable benefit/risk ratioapplicable to medical treatment. The type of disease of a patient,severity, activity of the drug, sensitivity to the drug, administrationtime, route of administration, excretion rate, treatment period, factorsincluding concomitant drugs, and other factors well-known in the medicalfield may determine the effective dose level.

The pharmaceutical composition, according to the present disclosure, maypreferably be administered simultaneously, separately, or sequentiallywith the concomitant drug in order to enhance the therapeutic effect andmay be administered by single or multiple doses. Considering all thefactors above, it is important to administer in an amount that mayderive the maximum effect with the minimum amount without side effects,which may be easily determined by those skilled in the art.Specifically, the effective amount of the pharmaceutical compositionaccording to the present disclosure may vary depending on the age, sex,condition, and weight of a patient, absorption of an active ingredientin vivo, inactivity rate and excretion rate, disease type, andconcomitant drugs.

The pharmaceutical composition of the present disclosure may beadministered to an individual by various routes. All modes ofadministration are predictable, including, for example, oraladministration, intranasal administration, transbronchialadministration, arterial injection, intravenous injection, subcutaneousinjection, intramuscular injection, or intraperitoneal injection.

The pharmaceutical composition of the present disclosure is determinedaccording to the type of drug that is the active ingredient, along withvarious related factors such as the disease to be treated, the route ofadministration, the age, sex, weight, and severity of the disease of apatient.

In another aspect of the present disclosure, the present disclosureprovides a method of inhibiting neuroinflammation, includingadministering the pharmaceutical composition to an individual. The term“individual” as used herein refers to a subject in need of treatment fora disease, and more specifically, a human or non-human primate andmammals such as a mouse, dog, cat, horse, and cow.

In addition, the pharmaceutical composition according to an exampleembodiment of the present disclosure may be a formulation selected fromthe group consisting of a solution, suspension, syrup, emulsion,liposome, acid, powder, granule, tablet, sustained-release agent, andcapsule.

Moreover, the composition may be a composition for oral administrationand have a formulation of a drug carrier or sustained-release agentincluding liposomes. In addition, the composition may be a compositionfor parenteral administration and have a formulation of a drug carrieror sustained-release agent including liposomes and ultrasound contrastagents.

The pharmaceutical composition of the present disclosure may beencapsulated in liposomes to provide stability in the formulation fordrug delivery. The liposomes used herein may be prepared by mixtures ofpolyols, surfactants, phospholipids, fatty acids, and water.

Polyols used in liposomes are not particularly limited and preferablyinclude propylene glycol, dipropylene glycol, 1,3-butylene glycol,glycerin, methylpropanediol, isoprene glycol, pentylene glycol,erythritol, xylitol, and sorbitol, but most preferably propylene glycol.

Any surfactant known in the art may be used to prepare liposomes, suchas anionic surfactants, cationic surfactants, amphoteric surfactants,and nonionic surfactants may be used, preferably anionic surfactants andnonionic surfactants are used. Specific examples of anionic surfactantsinclude alkyl acyl glutamate, alkyl phosphate, alkyl acetylate, di-alkylphosphate, and trialkyl phosphate. Specific examples of nonionicsurfactants include alkoxylated alkyl ethers, alkoxylated alkyl esters,alkyl poly glycosides, poly glyceryl esters, and sugar esters.

Phospholipids, another component used in the preparation of liposomes,are used as amphiphilic lipids, including natural phospholipids andsynthetic phospholipids, preferably lecithin. Fatty acids used inliposome preparation are high-grade fatty acids, preferably saturated orunsaturated fatty acids of the C12-22 alkyl chain, including, forexample, lauric acid, myristic acid, palmitic acid, stearic acid, oleicacid, and linoleic acid. The water used in the preparation of liposomesis generally deionized distilled water.

Preparation of liposomes may be performed by various methods known inthe art, but most preferably, the preparation is performed by applying amixture including the components to a high-pressure homogenizer. Theliposome system prepared thereby has the advantage of dissolving varioustypes of poorly soluble substances and stabilizing unstable substances,thereby maximizing drug delivery.

The pharmaceutical composition of the present disclosure may be preparedas a sustained-release agent to increase medication adherence byreducing the number of doses of the drug through continuous maintenanceof the effective blood concentration of the active ingredient.

Sustained-release agents are prepared by including sustained-releasecarriers and other adjuvants in addition to the active ingredients ofthe present disclosure. Various sustained-release carriers known in theart may be used for the sustained-release carriers that may be usedherein, but it is preferably polyethylene oxides.

In addition to other adjuvants, dilution carriers commonly used in thepharmaceutical field may be included. Examples of dilution carriers usedfor such purpose include lactose, dextrin, starch, microcrystallinecellulose, calcium mono hydrogen phosphate, calcium carbonate, sugarsand silicon dioxide, and other glidants such as zinc stearate ormagnesium to increase fluidity or other adjuvants available in thepharmaceutical field may be included.

The composition of the present disclosure may be for an administrationto a subject in which expression of peroxisome proliferator-activatedreceptor-γ coactivator-1α (PGC-1α) is reduced.

According to another aspect of the present disclosure, a healthfunctional food for ameliorating Parkinson's disease, including thecompound represented by Chemical Formula 1 as an active ingredient, maybe provided.

When the composition of the present disclosure is prepared as a foodcomposition or functional food composition, it may include not only thecompound of Chemical Formula I as an active ingredient but alsocomponents commonly added during food preparation, including, forexample, proteins, carbohydrates, fats, nutrients, seasonings, andflavoring agents. Examples of carbohydrates described above includemonosaccharides, such as glucose and fructose; disaccharides, such asmaltose, sucrose, and oligosaccharides; and polysaccharides, such asconventional sugars like dextrin and cyclodextrin, as well as sugaralcohols such as xylitol, sorbitol, and erythritol. As flavoring agents,natural flavoring agents and synthetic flavoring agents (saccharin,aspartame, etc.) may be used. In addition, when the food composition ofthe present disclosure is prepared as a drink, citric acid, liquidfructose, sugar, glucose, acetic acid, malic acid, fruit juice, Eucommiabark extract, jujube extract, licorice extract, and the like may befurther included in addition to the compound of Chemical Formula I ofthe present disclosure.

According to another aspect of the present disclosure, a method ofpreventing or treating Parkinson's disease, including administering, toa subject, a preparation including a compound represented by theChemical Formula 1 is provided.

MODES FOR CARRYING OUT THE INVENTION

Hereinafter, a preferred example embodiment of the present disclosurewill be described in detail with reference to the accompanying drawings.However, these example embodiments are intended only to illustrate thepresent disclosure, and the scope of the present disclosure will not beconstrued as being limited by these example embodiments.

Example 1: Preparation of a Reporter Cell Line and a Human Cell Line

In order to prepare lentiviruses for preparation of stable reporterSH-SY5Y cells (SH-PGC-1α-Luc), 1-kb pGL3-PGC-1α promoter-luciferase wascloned to pGreenFire (System Biosciences), and a lentivirus constructwas first prepared (provided by Akyosi Fukamizu from University ofTsukuba, Japan).

HEK293T cells cultured in 15 cm dish were subjected to transfection with32 μg of pGreenFire vector, 9 μg of VSVg envelope, 6.25 μg of Prev, and12.5 μg of pMDL.

After 48 hours, viral supernatants were collected and concentrated usingultracentrifugation, and viral pellets were dissolved with PBS. SH-SY5Ycells were treated with concentrated virus and screened with puromycin(1 μg/ml) a day later.

To culture human neuroblasts SH-SY5Y cells (ATCC, Manassas, Va.), 10%FBS (vol/vol, Welgene Gold Serum, cat #S 001-07) and DMEM (Welgene freshmedia DMEM, cat #LM 001-05) including antibiotics were used, wherein theculture was performed in an incubator (Forma Direct Heat CO2 incubator,Thermo Scientific) at 37° C. in the presence of 5% carbon dioxide.

Example 2: Compound Screening and Data Analysis to Identify Compoundsthat Increase the Expression of PGC-1α

The library of 8,320 compounds was sorted (Korean Chemical Bank,Daegeon) out of 230,000 drugs in consideration of the properties of thedrug, which is a combined library of 6,000 drugs and 2,320 spectrumcollections (Microsource, http://www.msdiscovery.com/spectrum.html)whose quality is controllable by liquid chromatography mass spectroscopy(LC-MS) (60% clinically approved drugs, 25% natural products, 15%bioactive drugs).

To determine the suitability of the experiment for high-throughputscreening (HTS), standard parameters including signal-to-background(S/B) ratio, daily rate of change, and rate of change for each plate, aswell as Z′ factor and coefficient of variation (CV) were measured.

All HTS experiments followed the NIH guidelines (High-ThroughputScreening Assay Guidance Criteria,http://www.ncats.nih.gov/research/reengineering/ncgc/assay/criteria/criteria.html).

SH-PGC-1α-Luc was dispensed in a white, flat-bottomed 96-well plate,where culture was performed to accommodate 10,000 cells in 100 μl ofDMEM (10% FBS+penicillin/streptomycin (P/S)) per well, and the cellswere stabilized in an incubator at 37° C. in the presence of 5% CO₂ for12 hours.

The next day, each drug was added to 50 μl of warmed DMEM to the finalconcentration of 20 μM, then 50 μl of DMEM was removed from thecell-containing plate, and 50 μl of drug-containing DMEM was added(final concentration of 10 μM). Cells were then exposed to the drug for48 hours, and luciferase activity was measured using SteadyGlo reagent(Promega).

Each plate had three internal controls, Daidzein (positive control, 10μM), and two negative controls (treated and untreated with DMSO). Theluciferase measurement values for each well are shown in the ratio basedon the average value of the untreated controls.

The Z′ factor of this experiment showed a value between 0.5 and 1. Thedifference between each well, plate, and experiment day was measured bycomparing the change in the control. DMSO resistance, stability of areagent, and experimental conditions were also checked to proveeffectiveness.

The activity measurement results for the PGC-1α promoter via luciferaseassay are shown in FIG. 4 . Luciferase activity was measured by readingthe first data, wherein the comparison was made based on DMSO, and theblue square box in FIG. 4 indicates drugs that are more than 1.5 timesactivated.

The results of the experiment performed with 14 drugs that increase theactivity of PGC-1α promoter by 2.5 times or more are shown in FIG. 5(YPD-01 in FIG. 5 is 2-(4-(1-hydroxypropan-2-yl)phenyl)isoindoline-1-onewhich is a compound represented by the Chemical Formula 1 herein, YPD-02is 2-[4-(3-oxo-1H-isoindol-2-yl)phenyl]propanoic acid, and YPD-03 isisopropyl 2-(4-(1-oxoisoindolin-2-yl)phenyl)propanoate).

Example 3: SH-SY5Y Cell and Tissue Sampling for Immunoblot

RIPA buffer was added to the SH-SY5Y cells and SN of C57L/6N mice,followed by homogenization with a homogenizer. Thereafter, the freezingand melting process was repeated three times, and quantification wasperformed using a BCA kit based on BSA to check the total amount ofprotein.

Lysates were identified by adding 2×SDS sample buffer, heating at 95° C.for 10 minutes to perform immunoblot, and then treating the desiredprotein with antibody.

In the immunoblot experiment, the darkness of the band was measuredusing the ImageJ (NIH, Bethesda, Mo., USA, http://rsb.info.nih.gov/ij/)program, and statistical analysis was performed using the darkness ofthe protein band in proportion to the loading control. The statisticalanalysis was performed using the GraphPad Prism version 7 (GraphPadSoftware) program. For the data, the unpaired two-tailed student'st-test was applied, wherein p<of 0.05 indicates statisticalsignificance.

In Example 4 below, when the sample shows a statistical significance bythe student's t-test compared to the control, it was marked with anasterisk (*p<0.05, ** p<0.01, and *** p<0.001).

Example 4: Effect of 2-(4-(1-Hydroxypropan-2-Yl)Phenyl)Isoindoline-1-Onein an SH-SY5Y Cell Line and the Mouse Brain

As shown in Example 3, an image of an immunoblot performed aftertreating the SH-SY5Y cell line with compounds (10 μM, 48 hours) is shownin FIG. 6 .

As shown in FIG. 6 , it was found that YPD-02(2-[4-(3-oxo-1H-isoindol-2-yl)phenyl]propanoic acid) and YPD-03(isopropyl 2-(4-(1-oxoisoindolin-2-yl)phenyl)propanoate) increased theamount of PGC-1α protein in accordance with an increase in the drugconcentration (0.01, 0.1, 1, 10 μM), while YPD-01(2-(4-(1-hydroxypropan-2-yl)phenyl)isoindolin-1-one) increased theamount of PGC-1α protein regardless of the concentration.

In FIG. 7 , an immunoblot is shown for protein derived from the SN ofmice fed with Chow, YPD-02(2-[4-(3-oxo-1H-isoindol-2-yl)phenyl]propanoic acid), or YPD-01(2-(4-(1-hydroxypropan-2-yl)phenyl)isoindoline-1-one) diet (0.5% w/w)for 1 week (quantitative graph quantified with β-actin/data wasexpressed as mean±SEM/statistical significance was measured by applyingthe unpaired two-tailed student t-test. *p<0.05, **p<0.01.).

From the results in FIG. 7 , it was found that YPD-01(2-(4-(1-hydroxypropan-2-yl)phenyl)isoindoline-1-one) successfullypassed through BBB and increased the protein level of PGC-1α in the SNof mice.

Example 5: Inhibitory Effect of YPD-01 on the Death of Dopamine Neuronsin Parkinson's Disease Model Mice

To prepare Parkinson's disease model mice, AAV-PARIS was stereotaxicallyinjected into the substantia nigra area. The mice were fed with Chow orYPD-01 (2-(4-(1-hydroxypropan-2-yl)phenyl)isoindoline-1-one) diet (0.5%w/w) for 4 weeks, and then the brain was collected and sectioned using amicrotome. Dopaminergic neurons were visualized by carrying out areaction in the mouse brain tissue sliced into 35 um sections using THantibody, a neuronal dopamine marker, then exposed to Vectastine ABC(Vector biolabs) and DAB (Sigma) solutions. Six rats were used in eachexperiment.

Introduction of Adeno-Associated Virus (AAV) Via Stereotaxic Injection

100 ul of pentobarbital (10 mg/ml) was injected into the abdomen of8-week-old mice for anesthetization, mouse pericranium was peeled off,and then the left brain (X: 1.2, Y: −3.2, Z: −4.5) and right brain (X:1.2, Y: 3.2, Z: −4.5) regions were marked based on the bregma. Themarked portion was drilled, and the virus was injected slowly (30seconds per 0.2 ul) through a syringe. After injecting the virus intothe left brain, followed by a 2-minute standby, the same procedure wasperformed on the other side. After the surgical suture, the mice werecarefully monitored, and breeding was followed in the cage until therecovery.

Immunoblot (Western Blot, WB)

Midbrains of mice into which AAV-PARIS was stereotaxically injected andwhich were fed with Chow or YPD-01 diet (0.5% w/w) for 4 weeks werecollected, and protein was extracted with RIPA lysis buffer. Theconcentration of protein was adjusted to 4 mg/ml by the BCA verificationmethod. Electrophoresis was performed in 7% polyacrylamide gel by mixingwith 2×Laemmli sample buffer. After transfer, the primary antibody andHRP-conjugated secondary antibody were attached, followed by developmentusing an ECL solution. AAV-GFP was used as a control of AAV-PARIS.

Immunohistochemistry

Mouse brain tissues sliced into 35 um ice sections were subjected to areaction with tyrosine hydroxylase (TH) antibody, a neuronal dopaminemarker, at 4° C. overnight, and a reaction was carried out the next dayusing biotin-conjugated secondary antibodies, followed by thedevelopment of the shape of dopamine neurons by exposing to VectastainABC (Vector Biolabs) and DAB (Sigma) solution. The developed braintissue was placed on a glass slide and examined under a microscope.

As shown in FIG. 8 , AAV-PAIS injection significantly killed dopamineneurons, and in mice fed with YPD-01 diet, the death of dopamine neuronswas meaningfully inhibited. Consistent with the result shown inimmunohistochemistry, as a result of performing immunoblot in thesubstantia nigra of mice using TH antibody, a dopamine neuronal marker(quantitative graph quantified with j-actin/data was expressed asmean±SEM/statistical significance was measured by applying one-wayANOVA. *p<0.05, ***p<0.001), it was re-identified that the death ofdopamine neurons by AAV-PARIS was inhibited by YPD-01 as confirmed bythe result found in immunohistochemistry.

Example 6: Inhibitory Effect on Behavioral Abnormality in Parkinson'sDisease by YPD01

In order to identify whether YPD-01 is effective in inhibiting the deathof dopamine neurons and Parkinson's disease-like behavioral abnormalityin the Parkinson's disease model, as shown in Example 5 above, a poletest, which is the most reliable behavioral experiment, was performed.

Pole Test

To investigate behavioral abnormality, Parkinson's disease model micewere transferred to a pole test cage to have the mice adapted for 3minutes, and then the mouse tails were held and placed on the tip of avertically erected pole. The time from the moment the mouse took off itshind foot from the tip of the pole until the mouse came down to thefloor was measured.

As shown in FIG. 9 , due to due to potential AAV-PARIS-induced death ofdopamine neurons, the time taken for mice injected with AAVPARIS to comedown from the top of the pole was about twice longer than that of miceinjected with AAV-GFP (used as a control). The symptoms of behavioralabnormality disappeared by YPD-01 intake, consistent with inhibition ofdeath of the dopamine neurons described in Example 5 above (data wasexpressed as mean±SEM/statistical significance was measured by applyingone-way ANOVA. *p<0.05).

Example 7: Verification that YPD-01 Increases Expression of PGC-1α inParkinson's Disease Model Mice

In the animal model experiment set up as in Example 5, whether theexpression of PGC-1α and the main target genes (NRF-1, Tfam) of PGC-1αincreases due to YPD-01 intake was measured by RT-qPCR method.

Reverse Transcription Quantitative Real-Time Polymerase Chain Reaction(RT-qPCR)

The midbrain of mice into which AAV-PARIS was stereotaxically injectedand which were fed with Chow or YPD-01 diet for 4 weeks was collected,and RNA was extracted using the total RNA extraction kit (IntronBiotechnology). cDNA was synthesized using the cDNA synthesis kit(Enzynomics) and oligo dT from the extracted RNA. Using primers and SYBRgreen (Qiagen) reagents for the gene to be analyzed, gene expression inRotor gene Q (Qiagen) was quantified via qPCR.

As identified in FIG. 10 , PARIS overexpression by AAV-PARIS causedsuppression of PGC-1α expression and reduced expression of its targetgenes. PARIS-induced suppression of expression of PGC-1α and main targetgenes (NRF-1, Tfam) of PGC-1α was significantly recovered in thesubstantia nigra of mice fed with YPD-01 (quantitative graph quantifiedwith β-actin/data was expressed as mean±SEM/statistical significance wasmeasured by applying one-way ANOVA. *p<0.05, **p<0.01; ns, notsignificant).

As a specific part of the present disclosure is described in detailabove, it will be apparent to those skilled in the art that suchspecific techniques are only preferred embodiments and the scope of thepresent disclosure is not limited thereby. Thus, the substantial scopeof the present disclosure will be defined by the appended claims andtheir equivalents.

1. A method of preventing or treating Parkinson's disease in a subjectin need thereof, comprising: administering a pharmaceutical compositioncomprising a compound represented by the following Chemical Formula 1

or a pharmaceutically acceptable salt thereof as an active ingredient tothe subject.
 2. The method of claim 1, wherein the pharmaceuticalcomposition increases expression of peroxisome proliferator-activatedreceptor-γ coactivator-1α (PGC-1α).
 3. The method of claim 1, whereinthe pharmaceutical composition has a formulation selected from the groupconsisting of a solution, suspension, syrup, emulsion, liposome, acid,powder, granule, tablet, sustained-release agent, and capsule.
 4. Themethod of claim 3, wherein the pharmaceutical composition is acomposition for oral administration and has a formulation of a drugcarrier or sustained-release agent comprising liposomes.
 5. The methodof claim 3, wherein the pharmaceutical composition is a composition forparenteral administration and has a formulation of a drug carrier orsustained-release agent comprising liposomes and ultrasonic contrastagents.
 6. The method of claim 1, wherein the pharmaceutical compositionis to be administered to a subject in which expression of peroxisomeproliferator-activated receptor-γ coactivator-1α (PGC-1α) is reduced. 7.A method of ameliorating Parkinson's disease to a subject in needthereof, comprising: administering a healthy functional food comprisinga compound represented by the following Chemical Formula 1

as an active ingredient to the subject.
 8. (canceled)